Tert-butyl alcohol, methanol, ethyl acetate, dimethyl sulfoxide (DMSO), 2,4,6-trinitrobenzene-1-sulfonic acid (TNBSA), N,N-dimethylacetamide (DMA), CuBr, 4-quinolinol, and N,N-diisopropylmethylamine (DIPEA) were purchased from Merck-Sigma Aldrich (Germany). Doxorubicin hydrochloride (DOX) was purchased from Meiji Seika, Japan. Dendritic cores; PFD-G4-TMP-azide (48 end groups), PFD-G3-TMP-azide (24 end groups), and PFD-G2-TMP-azide (12 end groups) dendrimers; PFD-G5-acetylene-ammonium (32 end groups); PFD-G4-acetylene-ammonium (16 end groups) and PFD-G3-acetylene-ammonium (8 end groups) dendrons were purchased from Polymer Factory (Sweden). PAMAM dendrimers were purchased from Dendritech (USA). Azo initiator V-70 was purchased from Wako Chemicals (Japan). p-SCN-Bn-Deferoxamine was purchased from Macrocyclics™ (USA). All solvents were of gradient grade or high-performance liquid chromatography (HPLC) grade.
P scn bn deferoxamine
Manufactured by Macrocyclics
Sourced in United States
P-SCN-Bn-deferoxamine is a chemical compound used in various laboratory applications. It serves as a bifunctional chelating agent, capable of binding to both metal ions and biomolecules. The compound's core function is to facilitate the coordination and detection of specific analytes in research and analytical settings.
Automatically generated - may contain errors
8 protocols using p scn bn deferoxamine
1
Dendrimer-Mediated Doxorubicin Delivery
2
Labeling P10-2 Fab with Deferoxamine
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Example 6
Subsequently, the present inventors labeled P10-2 Fab with a chelating agent mentioned above.
Specifically, a Fab fragment solution adjusted to 12.5 mg/mL with phosphate-buffered saline (pH 7.4) was adjusted to pH 9.0 by the addition of a 100 M sodium carbonate solution at 10 mM. p-SCN-Bn-deferoxamine (Macrocyclics, Inc.) was added thereto at a final concentration of 1 mM, and the resultant was reacted at 37° C. for 2 hours. p-SCN-Bn-deferoxamine has an isothiocyanate group and therefore reacts immediately with Lys of the Fab fragment. This was recovered through Amicon Ultra 10K-0.5 mL centrifugal filter to purify a chelating agent-labeled Fab fragment. This chelating agent-labeled P10-2 Fab was designated as P10-2 Fab DFO.
3
Radiolabeling Procedure for 68Ga-Deferoxamine
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All reagents were purchased from commercial sources as reagent or analytical grade and used without further purification. Solvents and chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA), Acros Organics (Geel, Belgium), AAPPTec (Louisville, KY, USA), or Fluorochem (Hadfield, UK). p-SCN-Bn-Deferoxamine was purchased from Macrocyclics (Plano, TX, USA). Anhydrous solvents were dried over 4 Å molecular sieves or stored as received from commercial suppliers. 68GaCl3 for radiolabeling was eluted from a 68Ge/68Ga-generator (Eckert & Ziegler Eurotope GmbH, Berlin, Germany) with 0.1 N HCl using a fractionated elution approach.
4
Dual-Modality Imaging of VCAM-1 Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To enable dual modality imaging, VCAM-1 or isotype control antibodies (immunoglobulin G, IgG; BD 553330, BD 553927) were buffer exchanged to PBS using NAP25 gel filtration tubes (GE Healthcare). Buffer exchanged antibodies were concentrated to 6 mg/kg (Amicon Ultra-4, 30 kDa, EMD Millipore) and 30% (volumetric) of 0.1 M sodium borate buffer pH 9.5 were added. The chelator p-SCN-Bn-Deferoxamine (Macrocyclics, B-705) was dissolved in DMSO, 4 mol deferoxamine/mol antibody were added to the antibody solution and incubated at 37°C for 90 minutes. Excess chelator was removed via buffer exchange and coupling efficiency was checked with LC–MS. Chelator coupled antibodies were covalently attached to tosylactivated Dynabeads (MPIO microparticles of iron oxide) following manufacturer's protocol (Invitrogen 65501) using 1 mg antibody for 25 mg Dynabeads. Binding of VCAM-MPIO and IgG-MPIO to stimulated endothelial cells was tested as outlined below. For MRI, antibody-MPIO were re-suspended 1 minute prior to injection using 0.45 mg particles for a 25 g mouse in 100 μl saline. For PET (positron emission tomography) imaging, antibody-MPIO were loaded with 68Ga at pH 4 for 15 minutes at room temperature followed by several washing steps leading to an activity of approximately 1200 μCi/mg particle.
5
Antibody Conjugation for 89Zr-Radiolabeling
6
Zr-89 Radiolabeled PLGA Nanoparticles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The conjugation of p-SCN-Bn-deferoxamine (DFO) to PLGA-NH2 nanoparticles and radiolabelling with 89Zr-oxalate (Korea Institute of Radiological and Medical Sciences, Seoul, Korea) was performed as previously described (Poot et al. 2019 (link)). Briefly, 5 mg of the synthesised PLGA-NH2 nanoparticles was thoroughly dispersed in 500 mL of PBS. Subsequently, 100 µg of amine-reactive p-SCN-Bn-deferoxamine (Macrocyclics, TX, USA) in dimethyl sulfoxide (2 mg/mL) was added to the nanoparticles, thoroughly mixed, and the pH was adjusted to 9.0 using 1.0 M Na2CO3 solution. The thiocarbamide bond reaction was allowed to proceed at 37 °C for 1 h at 550 rpm using a thermomixer. The DFO-conjugated PLGA-NH2 nanoparticles were collected via centrifugation at 3500 rpm for 5 min and washed thrice with PBS to remove unreacted p-SCN-Bn-deferoxamine. For 89Zr radiolabelling, 35–40 MBq of 89Zr-oxalate was diluted in 200 µL of 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid buffer (0.5 M), neutralised with 1.0 M Na2CO3. The 89Zr solution was mixed with the DFO-conjugated nanoparticles and radiolabelled at room temperature for 1 h at 350 rpm. The radiolabelled PLGA-NH2 nanoparticles were washed thrice with PBS, and the radiochemical yield was determined using a Bioscan AR-2000 radio-thin layer chromatography scanner (Eckert & Ziegler, Berlin, Germany).
7
Labeling Fab Fragment with Deferoxamine
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Example 6
Subsequently, the present inventors labeled P10-2 Fab with a chelating agent mentioned above.
Specifically, a Fab fragment solution adjusted to 12.5 mg/mL with phosphate-buffered saline (pH 7.4) was adjusted to pH 9.0 by the addition of a 100 M sodium carbonate solution at 10 mM. p-SCN-Bn-deferoxamine (Macrocyclics, Inc.) was added thereto at a final concentration of 1 mM, and the resultant was reacted at 37° C. for 2 hours. p-SCN-Bn-deferoxamine has an isothiocyanate group and therefore reacts immediately with Lys of the Fab fragment. This was recovered through Amicon Ultra 10K-0.5 mL centrifugal filter to purify a chelating agent-labeled Fab fragment. This chelating agent-labeled P10-2 Fab was designated as P10-2 Fab DFO.
8
Labeling P10-2 Fab with Deferoxamine
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Example 6
Subsequently, the present inventors labeled P10-2 Fab with a chelating agent mentioned above.
Specifically, a Fab fragment solution adjusted to 12.5 mg/mL with phosphate-buffered saline (pH 7.4) was adjusted to pH 9.0 by the addition of a 100 M sodium carbonate solution at 10 mM. p-SCN-Bn-deferoxamine (Macrocyclics, Inc.) was added thereto at a final concentration of 1 mM, and the resultant was reacted at 37° C. for 2 hours. p-SCN-Bn-deferoxamine has an isothiocyanate group and therefore reacts immediately with Lys of the Fab fragment. This was recovered through Amicon Ultra 10K-0.5 mL centrifugal filter to purify a chelating agent-labeled Fab fragment. This chelating agent-labeled P10-2 Fab was designated as P10-2 Fab DFO.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!